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Biomedical subjects

G B Anderson

Publications and source records attributed to G B Anderson.

At least 19 recordsLinked to original sources
PubMed2006

Effect of porcine zonae pellucidae immunisation on ovarian follicular development and endocrine function in domestic ewes (Ovis aries).

Domestic ewes (Ovis aries) were immunised with porcine zonae pellucidae (pZP) or pZP conjugated to keyhole limpet haemocyanin (KLH) in adjuvant(s) to examine the feasibility of the species to serve as a model for further development of pZP-based vaccines in ungulates. Two immunisation groups were employed, with a third group receiving only adjuvant (n = 5 per group). Early in the study, oestrous activity was monitored by the use of a vasectomised ram fitted with a marking harness. Eventually, ewes were exposed to an intact ram for breeding. In addition, weekly serum and every-other-day faecal samples were collected to measure pZP antibodies and progesterone metabolite concentrations respectively. At the conclusion of the study, fecundity was established, and ovarian tissue was examined. Ewes immunised against pZP : KLH with adjuvant produced minimal antibody absorbance levels, displayed normal oestrous cycles, became pregnant upon introduction of the intact ram and exhibited normal ovarian histopathology. Ewes immunised against pZP with adjuvant produced high antibody absorbance levels, were acyclic following primary immunisation and were infertile. Examination of the ovarian tissue revealed atrophic changes that included: (1) the absence of growing follicles; (2) significant reduction in the number of primordial follicles; and (3) the presence of abnormal granulosa cell clusters lacking oocytes. Antisera displayed immunoreactivity to the major components of pZP, and immunohistochemical labelling of ovarian tissue showed specificity to the ZP. These data are the first generated in an ungulate species showing deleterious effects of pZP immunisation on folliculogenesis and oestrous cyclicity.

Adjuvants, Immunologic↗
PubMed2006

Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland.

The potential for applying biotechnology to benefit animal agriculture and food production has long been speculated. The addition of human milk components with intrinsic antimicrobial activity and positive charge to livestock milk by genetic engineering has the potential to benefit animal health, as well as food safety and production. We generated one line of transgenic goats as a model for the dairy cow designed to express human lysozyme in the mammary gland. Here we report the characterization of the milk from 5 transgenic females of this line expressing human lysozyme in their milk at 270 microg/mL or 68% of the level found in human milk. Milk from transgenic animals had a lower somatic cell count, but the overall component composition of the milk and milk production were not different from controls. Milk from transgenic animals had a shorter rennet clotting time and increased curd strength. Milk of such nature may be of benefit to the producer by influencing udder health and milk processing.

Animals↗
PubMed2004

Evidence of increased substrate availability to in vitro-derived bovine foetuses and association with accelerated conceptus growth.

Changes in placental development have been associated with foetal abnormalities after in vitro embryo manipulations. This study was designed to investigate bovine conceptus development and substrate levels in plasma and fluids in in vivo- and in vitro-produced (IVP) concepti and neonates. In vivo-produced and IVP embryos were derived by established embryo production procedures. Pregnant animals from both groups were slaughtered on days 90 or 180 of gestation, or allowed to go to term. Conceptus and neonatal physical traits were recorded; foetal, maternal and neonatal blood, and foetal fluids were collected for the determination of blood and fluid chemistry, and glucose, fructose and lactate concentrations. Placental transcripts for specific glucose transporters were determined by quantitative RT-PCR. No significant differences in uterine and conceptus traits were observed between groups on day 90. On day 180, larger uterine, placental and foetal weights, and an increase in placental gross surface area (SA) in IVP pregnancies were associated with increased glucose and fructose accumulation in foetal plasma and associated fluids, with no differences in the expression of components of the glucose transporter system. Therefore, the enlarged placental SA in IVP pregnancies suggests an increase in substrate uptake and transport capacity. Newborn IVP calves displayed higher birth weights and plasma fructose concentrations soon after birth, findings which appeared to be associated with clinical and metabolic distress. Our results indicated larger concepti and increased placental fructogenic capacity in mid- to late IVP pregnancies, features which appeared to be associated with an enhanced substrate supply, potentially glucose, to the conceptus.

Animals↗
PubMed2004

Hot topic: using a stearoyl-CoA desaturase transgene to alter milk fatty acid composition.

Stearoyl-CoA desaturase enzyme converts specific medium- and long-chain saturated fatty acids to their monounsaturated form. Transgenic goats expressing a bovine beta-lactoglobulin promoter-rat stearoyl-CoA desaturase cDNA construct in mammary gland epithelial cells were produced by pronuclear microinjection. The fatty acid composition of milk from 4 female transgenic founders was analyzed on d 7, 14, and 30 of their first lactation. In 2 animals, the expression of the transgene changed the overall fatty acid composition of the resulting milk fat to a less saturated and more monounsaturated fatty acid profile at d 7 of lactation; however, this effect diminished by d 30. In addition, one animal had an increased proportion of the rumen-derived monounsaturated fatty acid C18:1 trans11 converted by stearoyl-CoA desaturase to the conjugated linoleic acid isomer C18:2 cis9 trans11. Milk that has higher proportions of monounsaturated fatty acids and conjugated linoleic acid may have benefits for human cardiovascular health.

Animals↗
PubMed2002

The placenta as a contributor to production of large calves.

Unusually high birth weights frequently occur in calves born from cultured bovine embryos. The mechanism(s) through which in vitro manipulations during early cleavage are translated to enhanced fetal growth is (are) incompletely understood. Accelerated growth is primarily prenatal, and the placenta of an in vitro-derived conceptus could account for abnormal fetal growth. Results from a systematic comparison of placental morphology and function in bovine concepti produced in vitro versus in vivo are discussed.

Animals↗
PubMed2001

Evidence against humoral immune attack as the cause of sheep-goat interspecies and hybrid pregnancy failure in the doe.

The failure of interspecies and hybrid pregnancies between the domestic sheep (Ovis aries) and goat (Capra hircus) is not completely understood. The sheep-goat hematopoietic chimera is a unique model for studying the role of the maternal immune response in failure of interspecies and hybrid pregnancies between these species. Hematopoietic chimeras were created by in utero transplantation of sheep fetal liver cells into goat fetuses. The resulting chimeric females were recipients of sheep demi-embryos genetically identical to their sheep cells and/or were bred to a ram to create a hybrid pregnancy. Pregnancy sera were analyzed for the presence of anti-species antibodies (Ab) using a lymphocyte microcytotoxicity assay. None of the concepti survived to term. Gross and histological evaluations of two interspecies sheep concepti revealed abnormal placentome formation. The humoral immune response of several hematopoietic chimeras to the challenging concepti differed from control animals. We observed delayed onset of Ab production, low and absent titers, and persistent Ab titers with delayed fetal death. Ultrasonography typically revealed normal fetal development associated with high volumes of placental fluids and retarded placentome development. We conclude that fetal death was associated with abnormal placental development that was not the result of maternal humoral immune attack.

Animals↗
PubMed2001

Fetal leukocyte trafficking as a stimulus for the production of maternal antibodies in the goat.

The production of antibodies during pregnancy or after parturition is a natural occurrence in many mammalian species. Fetal cells have been detected in the peripheral blood of women and mice and are thought to be the immune stimulus for antibody production. The aim of this study was to investigate if the production of maternal anti-fetal antibodies during ruminant pregnancy is the result of fetal leukocyte trafficking across the placenta. Maternal pregnancy serum was collected from 94 does whose fetuses received sheep hematopoietic stem cells via in utero transplantation at 49 to 62 d of gestation. Serum samples were collected before surgery and at weekly intervals throughout gestation. A lymphocyte microcytotoxicity assay was used to screen the serum samples from does that carried chimeric fetuses to term (n = 75). Of these 75 does, 28 parous does had presurgery serum that contained alloreactive antibodies. Nine of the 75 does had nonreactive presurgery serum, but they produced alloreactive antibody titers during gestation. Xenoreactive antibodies were detected in the pregnancy sera from 2 of the 75 does tested. Hemolytic assays confirmed the species-specificity of the xenoreactive serum from these 2 does. In view of the fact that hematopoietic cells were the only source of anti-sheep antibody stimulation in this model, we propose that fetal leukocyte trafficking does take place across the caprine placenta.

Animals↗
PubMed2001

Maternal-conceptus signalling during early pregnancy in mares: oestrogen and insulin-like growth factor I.

Embryonic production of oestrogen is thought to play an important role in conceptus-maternal signalling during early pregnancy in mares, and may be regulated in an autocrine or paracrine fashion by insulin-like growth factor I (IGF-I). In this study, the hypothesis that IGF-I stimulates embryonic oestrogen synthesis, which in turn stimulates uterine IGF-I secretion was tested. Specific sources of IGF-I in the uterine lumen were characterized. Preimplantation embryos, uterine biopsies, and uterine flush fluids were collected on day 13 of pregnancy. Embryos were cultured whole for 24 h, or dispersed and incubated in serum-free culture medium supplemented with androstenedione or testosterone (0-10 microg ml(-1)) and IGF-I (0-100 microg ml(-1)). Oestrogen synthesis was increased by addition of androgen, but there was no dose-dependent effect of IGF-I. Endometrial explants were cultured for 24, 48 and 72 h in serum-free medium supplemented with oestradiol. IGF-I was measured by radioimmunoassay in embryo-conditioned medium, explant culture medium, blastocoelic fluid, concentrated (x 100) uterine flush fluid and endometrial-tissue homogenate. Both the embryo and endometrium produced significant quantities of IGF-I, indicating a role for this growth factor in autocrine-paracrine signalling during early pregnancy. However, secretion of IGF-I by endometrial explants was not modulated by oestrogen.

Animals↗
PubMed2001

Contributions of tissues to expression of a major gene for growth in chimeric mice.

The objective of this research was to determine the effect of genotype of particular tissues and organs on the growth of a chimera produced by aggregating an embryo homozygous for the high-growth (hg) allele with an embryo from a control line of wild-type mice lacking the deletion responsible for the high-growth phenotype. A total of 107 chimeric mice were produced, 81 males and 26 females. The preponderance of male chimeras closely fit the expected 3:1 ratio of males to females in balanced chimeric populations, suggesting that neither of the two genetic lines appeared to have a selective advantage for contributions to chimeric tissues. No single correlation between individual tissue chimerism and mouse growth was remarkable relative to correlations of chimerism of other tissues to growth. Other statistical models, including multiple regression techniques, did nothing to identify particular organs, tissues or various combinations that improved the explanation of expression of the high-growth phenotype by percentage of high-growth cells in that organ. These observations suggest that the chromosomal deletion thought to cause the high-growth phenotype has a systemic effect, without dominant effects of or on individual organs or tissues; the precise mechanism has yet to be determined.

Animals↗
PubMed2000

Successful pregnancy in goats carrying their genetically identical conceptus.

Mammalian pregnancies are naturally allogeneic, but syngeneic pregnancies have been carried to term in laboratory animal species. The need for maternal immune recognition during mammalian pregnancy is still unclear. Allogeneic pregnancies are protected from maternal immune attack by the nature of the trophoblast and its interactions with maternal tissues at the maternal-fetal interface. Syngeneic pregnancy models and the success of pregnancies in immunosuppressed mice challenge the necessity of a maternal immune response in mammals. This study was designed to investigate if outbred, domestic sheep and goats can successfully establish and maintain a syngeneic pregnancy. Embryo splitting and cryopreservation techniques were used to enable sheep and goat demi-embryos to be transferred to genetically identical females. Allogeneic pregnancies were established from the transfer of demi-embryos subjected to the same manipulations to assess demi-embryo survival and pregnancy rates under conventional immune compatibility conditions. Syngeneic pregnancies were established and carried to term in goats (2/11) but not in sheep (0/24). Microsatellite and DNA fingerprinting analyses confirmed that each kid was a genetically identical twin to the female that carried it to term. Our results demonstrated that genetic disparity is not required for the establishment and maintenance of pregnancy in goats, but our results were inconclusive for sheep.

Animals↗
PubMed2000

Tissue-specific localization of cytochrome P450 aromatase in the equine embryo by in situ hybridization and immunocytochemistry.

Estrogen production by the preimplantation equine embryo is presumed to be important in maternal-conceptus communication in the mare. The synthesis of C(18) estrogens from C(19) androgens requires cytochrome P450 aromatase (P450(arom)) in the conceptus, but little information is available on the specific tissue location or potential developmental patterns of expression for the horse. The goal of this research was to localize P450(arom) in the equine conceptus by immunocytochemistry and in situ hybridization. Intact blastocyst-stage embryos were collected by nonsurgical flush on Days 12-15 of pregnancy, fixed in 4% paraformaldehyde, and paraffin-embedded. Aromatase protein was localized using rabbit anti-human placental aromatase antiserum with a detection system utilizing peroxidase and 3-amino-9-ethylcarbazole. For in situ hybridization, tissue sections were incubated with sense or antisense [(35)S]UTP-labeled cRNA probes prepared from equine aromatase cDNA. Aromatase protein and transcript were abundant in the extraembryonic trophectoderm but absent from embryonic ectoderm. No P450(arom) expression was detected in abembryonic endoderm or mesoderm. Aromatase expression was demonstrated in the endoderm beneath the disc (hypoblast). This pattern of P450(arom) expression in the equine blastocyst closely resembles that seen transiently in the porcine embryo, suggesting that regulatory mechanisms conferring tissue specificity may be conserved.

Animals↗
PubMed2000

CT angiography for the detection and characterization of carotid artery bifurcation disease.

BACKGROUND AND PURPOSE: Computed tomographic angiography (CTA) is a relatively new and minimally invasive method of imaging intracranial and extracranial blood vessels. The main purpose of this study was to compare CTA to the current gold standard of arterial imaging, digital subtraction angiography (DSA), for the detection and quantification of carotid artery bifurcation stenosis. We also compared Doppler ultrasound (US) with these 2 techniques. METHODS: In a prospective study, 40 patients (80 carotid arteries) underwent CTA, US, and DSA. Patients chosen for inclusion were symptomatic with TIAs or stroke and had initial US screening that indicated >50% carotid stenosis on the side appropriate for the symptoms. Source axial, maximum intensity projection (MIP), and shaded-surface display (SSD) images were produced for each CTA study. The US, CTA, and DSA images were reviewed, with the degree of stenosis quantified and presence of ulcers determined; each type of imaging was reviewed by a separate investigator blinded to the results of the other 2 modalities. The results of CTA and US imaging were compared with the DSA images for degrees of carotid stenosis. RESULTS: CTA source axial images correlated with DSA more closely than MIP or SSD images for all degrees of stenosis. The correlation between US and DSA (0.808) was poorer than that between CTA and DSA (0.892 to 0.922). CTA performed well in the detection of mild (0% to 29%) carotid stenosis, as well as carotid occlusion, with values for sensitivity, specificity, and accuracy near 100%. In determining that a stenosis was >50% by DSA measurement, CTA was again useful, with a sensitivity, specificity, and accuracy of 89%, 91%, and 90%, respectively. While CTA was quite specific and accurate in identifying degrees of stenoses in either the 50% to 69% or the 70% to 99% ranges, in this task it was much less sensitive: 65% for 50%-69% stenosis and 73% for 70%-99% stenosis. These results did not change significantly when only the data from the most clinically relevant symptomatic arteries were analyzed. CTA was found to correlate quite well with DSA in the detection of ulcers associated with the carotid stenosis. CONCLUSIONS: CTA was found to be an excellent examination for the detection of carotid occlusion and categorization of stenosis in either the 0%-29% or >50% ranges. However, CTA was unable to reliably distinguish between moderate (50%-69%) and severe (70%-99%) stenosis, which is an important limitation in the investigation and treatment of carotid stenosis.

Aged↗
PubMed1999

Establishment of pluripotent cell lines from porcine preimplantation embryos.

Embryonic stem (ES) cells are pluripotent cells isolated from in vitro culture of preimplantation embryos. Experiments were undertaken to identify preimplantation embryonic stages and culture conditions under which pluripotent, porcine embryo-derived cell lines could be isolated. Cell lines were established from in vitro culture of intact, porcine early hatched blastocysts and isolated inner cell masses (ICM) from intermediate and late hatched blastocysts on feeder layers prepared from permanent mouse embryonic fibroblasts (STO). The cells of these porcine embryo-derived cell lines had a morphology similar to that of murine ES cells, but colony morphology was more epithelial-like. The cell lines retained a normal diploid karyotype, consistently expressed alkaline phosphatase activity, and survived cryopreservation. When subjected to in vitro differentiation, either spontaneous or induced, the embryo-derived cell lines differentiated extensively into a wide range of cell types representing the 3 embryonic germ layers. In vivo pluripotency of the cells was demonstrated by birth of a chimeric piglet, documented by pigmentation and DNA markers, and the ability to direct the development of nuclear-transfer embryos to the blastocyst stage. Such pluripotent embryo-derived cells provide a potential route for porcine genetic manipulation.

Alkaline Phosphatase↗
PubMed1999

Progestin implants can rescue demi-embryo pregnancies in goats: a case study.

Survival after transfer of demi-embryos (i.e., half-embryos produced by embryo splitting) to recipients usually is lower than survival after transfer of intact embryos. Reduced survival after demi-embryo transfer could be due to loss of viability after splitting, failure of a viable demi-embryo to prevent corpus luteum (CL) regression in the recipient female, or a combination of factors. From a retrospective analysis of pregnancy and embryo survival rates after demi-embryo transfer in sheep and goats, we report the rescue of caprine demi-embryo pregnancies in which CL regression occurred at the end of diestrus despite the presence of a viable conceptus in the uterus with progestin implants. Day 5 or 6 morulae and blastocysts were flushed from superovulated ewes and does and split into demi-embryos of approximately equal halves. Demi-embryos were either transferred fresh to synchronized recipients of the homologous species or frozen in liquid nitrogen. Approximately half of the recipient does and ewes were treated with norgestomet implants on Day 10 of the embryo transfer cycle and again 2 wk later. Serum collected on Day 25 from recipients with implants was assayed for progesterone to determine if a CL of pregnancy had been maintained. Pregnancy was diagnosed by ultrasonography on Day 35 of gestation. Corpus luteum regression occurred despite the presence of a viable conceptus in the uterus in 6 of 55 progestin-treated caprine demi-embryo recipients and in 0 of 66 ovine demi-embryo recipients. Five of the caprine pregnancies were maintained to term with norgestomet implants and produced 5 live kids. The sixth fetus, which was carried by a progestin implant-treated 8-mo-old doeling, died at approximately 50 d of gestation. These results suggest that, at least in goats, some demi-embryos may provide inadequate signaling for maternal recognition of pregnancy, and such pregnancies can be rescued with progestin treatment to the doe.

Animals↗
PubMed1999

Infertility in a line of mice with the high growth mutation is due to luteal insufficiency resulting from disruption at the hypothalamic-pituitary axis.

Animals with extreme body growth frequently experience poor reproductive performance, but the cause for this association has not been clearly established. A line of mice homozygous for the high growth (hg) mutation, a mutation that has a major effect on post-weaning growth, exhibits several reproductive deficits including an abnormally high incidence of luteal failure. The objective of the present study was to investigate luteal failure in high-growth (HG) mice during pregnancy and to determine whether the cause of the apparent luteal failure resides in the ovary or the hypothalamic-pituitary unit. In HG females with a demonstrated history of infertility, progesterone injections (1 mg s.c. daily) beginning on Day 1 postcoitum (p.c.) increased the proportion of animals pregnant at Day 17 of gestation. Twice-daily injections of 100 microgram of ovine prolactin (PRL) in alkaline saline given to another group of females beginning on Day 1 p.c. increased the proportion of HG females that were pregnant on Day 6 of gestation compared with saline-injected HG females, although PRL did not increase the pregnancy rate in HG females when compared with a group of noninjected control females. When ovaries from HG females were transplanted into ovariectomized congenic C57 hosts, the C57 graft hosts displayed normal estrous cycles, and upon mating the majority of graft hosts became pregnant. In contrast, when ovaries from C57 females were transplanted into ovariectomized HG hosts, the HG graft hosts displayed normal estrous cycles, but upon mating most were unable to maintain pregnancy. These results suggest that the hypothalamic-pituitary unit of the HG female provides an inadequate signal to the ovaries to maintain pregnancy. Luteal failure in HG females may be due to insufficient PRL as required for establishment and early maintenance of the CL during pregnancy in mice.

Animals↗
PubMed1999

Computed tomographic angiography versus digital subtraction angiography for the diagnosis and early treatment of ruptured intracranial aneurysms.

OBJECTIVE: Computed tomographic angiography (CTA) is a rapid and minimally invasive method of detecting intracranial aneurysms. We wished to determine whether CTA could replace digital subtraction angiography (DSA) in the diagnosis and operative planning of ruptured cerebral aneurysms. METHODS: In a prospective study, patients with subarachnoid hemorrhage diagnosed by plain computed tomography underwent CTA, DSA, or both. Computed tomographic scans and CTA studies were first reviewed by the treating surgeon, along with a neuroradiologist, and a decision to proceed to DSA or directly to surgery was made on the basis of the type and quality of information provided by CTA. All patients underwent postoperative DSA. RESULTS: A total of 173 patients were studied. In 24 patients, both CTA and DSA were negative for a source of subarachnoid hemorrhage. Twelve patients underwent DSA without prior CTA because a technologist capable of performing CTA was not available when the patient was evaluated. Nine patients in poor neurological condition underwent CTA, and all tested positive for aneurysms but died without surgical intervention. Of the 126 patients who underwent CTA and surgery, 65 (52%) also required preoperative DSA. The decision to proceed to DSA after CTA was influenced by aneurysm location; posterior communicating artery (62%) and posterior circulation locations (67-75%) more commonly proceeded to DSA than middle cerebral artery aneurysms (34%; 0.025 > P > 0.01). The sensitivity and specificity of CTA for the detection of all aneurysms, ruptured and unruptured, in the group of patients who underwent both types of angiograms preoperatively were 84 and 100%, respectively. In the group of 61 patients in whom aneurysm surgery was performed on the basis of CTA results alone, the sensitivity and specificity for the detection of all aneurysms, as compared with postoperative DSA, were 90 and 100%, respectively. Missed aneurysms (n = 24) were always small (<4 mm) and were usually found in patients with multiple aneurysms in whom the larger, ruptured aneurysm was identified by CTA. In one patient, the aneurysm missed by preoperative CTA would have resulted in a different operation if detected preoperatively. CONCLUSION: It is possible to proceed to ruptured aneurysm repair entirely on the basis of good-quality CTA studies that demonstrate an aneurysm consistent with the pattern of bleeding observed on plain computed tomography (48% of the patients in this series and most common middle cerebral artery aneurysms). However, detection of small unruptured aneurysms in patients with multiple lesions remains a problem.

Adult↗
PubMed1999

Short communication: lack of stage-specific embryonic antigen-1 expression by bovine embryos and primordial germ cells.

The objective of this study was to determine whether stage-specific embryonic antigen-1, a cellular marker commonly used to identify murine undifferentiated embryonic cells, is also a useful marker for bovine pluripotent cells. Expression of stage-specific embryonic antigen-1 was examined by indirect immunohistochemistry on bovine preimplantation embryos and on primordial germ cells contained in the genital ridge. Expression of stage-specific embryonic antigen-1 was not observed in any of the cleavage-stage bovine embryos examined, including one-cell, two-cell, four-cell, eight-cell, morula, and blastocyst stages, nor in tissue sections of bovine genital ridges collected from embryos on d 34, 37, and 40 of gestation. As expected, expression of stage-specific embryonic antigen-1 was detected on murine preimplantation embryos and on murine teratocarcinoma cells. Results of this study indicate that, unlike in the mouse, stage-specific embryonic antigen-1 is not a useful cellular marker for pluripotent bovine embryonic cells or bovine primordial germ cells.

Animals↗